Regulation of Hypoxia-inducible Factor 1 by Cyclin Dependent Kinases

细胞周期依赖性激酶对缺氧诱导因子 1 的调节

• 批准号: 8456396
• 负责人: Noel Andrew Warfel
• 金额: $ 4.39万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2013-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8456396
• 关键词: Accounting; Acute; Address; Automobile Driving; Biochemical; Biological; CDC2 Protein Kinase; CDK4 gene; Cell Death; Cessation of life; Clinical; Cyclin-Dependent Kinase Inhibitor; Cyclin-Dependent Kinases; Data; Development; Diagnosis; Genetic Transcription; Goals; Hypoxia; Imaging technology; Kidney Neoplasms; Laboratory Study; Mass Spectrum Analysis; Measures; Mediating; Mediator of activation protein; Modeling; Molecular; Monitor; Mutation; Neoplasm Metastasis; Oxygen; Patients; Phosphotransferases; Physiology; Population; Post-Translational Protein Processing; Proteins; Regulation; Renal Cell Carcinoma; Renal carcinoma; Reporter; Repression; Research; Resistance; Solid Neoplasm; System; Testing; Time; Translations; Tumor Angiogenesis; Tumor Burden; United States; Von Hippel-Lindau Syndrome; angiogenesis; cancer therapy; deprivation; disorder control; driving force; glucose metabolism; hypoxia inducible factor 1; improved; in vitro Assay; in vivo; new therapeutic target; novel; novel strategies; novel therapeutic intervention; outcome forecast; patient population; pre-clinical; public health relevance; response; therapeutic angiogenesis; therapy resistant; transcription factor; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor progression

DESCRIPTION (provided by applicant): The hypoxia-inducible factor 1 (HIF-1) transcription factor is a critical mediator of the cellular response to oxygen deprivation. The ? subunits of HIF-1 are rapidly degraded in the presence of oxygen, but are stabilized under hypoxic conditions and activate the transcription of genes that promote angiogenesis, survival, invasion, and glucose metabolism. HIF-1 can be activated by genetic alterations that enhance the transcription or inhibit the degradation of HIF-1?. For example, von Hippel-Lindau (VHL) disease predisposes sufferers to aggressive tumors of the renal system, which are characterized by the stabilization of HIF-1?. The constitutive expression of HIF-1? drives the development of these tumors, which are highly angiogenic and resistant to therapy. Thus, HIF-1 inhibition represents a promising strategy for the treatment of patients with hypoxic and VHL-deficient tumors. While the oxygen-dependent regulation of HIF-1? has been well studied, the oxygen- independent regulation of HIF-1? is not completely understood. We have evidence that the stability of HIF-1? is regulated by cyclin dependent kinases (CDKs), specifically CDK1 and CDK4. The central hypothesis driving this research is that HIF-1? expression is regulated by CDKs, and identification of the molecular mechanisms underlying this regulation will produce novel strategies to inhibit tumor survival, angiogenesis, and therapeutic resistance associated with HIF-1 activation. We will address this hypothesis via the following aims: 1] Identify the mechanism by which CDKs regulate HIF-1? expression. 2] Test the anti-tumor efficacy of CDK inhibition on HIF-1? expressing tumors, and identify synergistic combinations with approved anticancer therapies. This proposal relies on three major approaches: 1] monitoring the expression levels and activity of HIF- 1 in response to CDK inhibition, 2] the use of mass spectrometry to identify of novel post-translational modifications that alter HIF-1? stability, and 3] utilizing CDK inhibitors in HIF-1? expressing tumors in combination with standard therapies. Preliminary data indicate that inhibition of CDK kinase activity significantly decreases the stability of HIF-1? at the protein level. Therefore, we plan to perform mass spectrometry and biochemical analyses to identify novel post-translational modifications on HIF-1? and determine the mechanism through which CDKs regulate HIF-1a expression. Next, we will investigate the efficacy of CDK inhibition as a novel therapeutic approach to target renal tumors that constitutively express HIF-1? due to the loss of VHL. First, in vitro assays will be used to assess the efficacy of CDK inhibitors and identify promising combinations with standard anticancer therapies. Then, in vivo tumor models will serve to determine if CDK-mediated repression of HIF-1? inhibits tumor growth and angiogenesis, and sensitizes HIF-1?-driven tumors to undergo cell death in response to standard therapies. Ultimately, the goal of this proposal is to facilitate the clinical translation of CDK inhibitors for use in tumors with constitutively active IF-1 while also yielding biological information about how HIF-1? stability is regulated by CDKs.

描述(由申请人提供)：缺氧诱导因子1(HIF-1)转录因子是细胞对缺氧反应的关键介质。那个？HIF-1的亚基在有氧条件下迅速降解，但在低氧条件下稳定下来，并激活促进血管生成、存活、侵袭和葡萄糖代谢的基因的转录。HIF-1可被增强转录或抑制HIF-1降解的基因改变所激活。例如，von Hippel-Lindau(VHL)病易患以HIF-1稳定为特征的肾脏侵袭性肿瘤。HIF-1？推动这些肿瘤的发展，这些肿瘤具有高度的血管生成性和对治疗的抵抗力。因此，抑制HIF-1是治疗缺氧性和VHL缺陷性肿瘤的一种有前途的策略。而HIF-1的氧依赖调节？已有的研究表明，HIF-1的氧非依赖性调节？还没有完全被理解。我们有证据表明HIF-1的稳定性？受细胞周期蛋白依赖性激酶(CDKs)的调节，特别是CDK1和CDK4。推动这项研究的中心假设是HIF-1？CDK的表达受CDKs的调控，识别这种调控的分子机制将产生抑制肿瘤存活、血管生成和与HIF-1激活相关的治疗耐药的新策略。我们将通过以下目标解决这一假说：1)确定CDK调节HIF-1的机制？表情。2)检测CDK抑制对HIF-1的抗肿瘤作用。表达肿瘤，并确定与已批准的抗癌治疗的协同组合。这一建议依赖于三个主要方法：1)监测CDK抑制反应中HIF-1的表达水平和活性，2)使用质谱学来确定改变HIF-1的新的翻译后修饰？稳定性，以及 3)在HIF-1中应用CDK抑制剂？在结合标准治疗的情况下表达肿瘤。初步数据表明，抑制CDK活性显著降低了HIF-1？在蛋白质水平上。因此，我们计划进行质谱和生化分析，以确定新的翻译后修饰的HIF-1？并确定CDKs调控HIF-1a表达的机制。接下来，我们将研究抑制CDK作为一种新的治疗方法对靶向表达HIF-1？由于VHL的损失。首先，将使用体外试验来评估 CDK抑制剂的有效性，并确定与标准抗癌治疗的有希望的组合。那么，体内肿瘤模型将用于确定CDK是否介导对HIF-1的抑制？抑制肿瘤生长和血管生成，并使HIF-1？驱动的肿瘤对标准治疗敏感，使其经历细胞死亡。最终，这项建议的目标是促进CDK抑制剂的临床翻译，用于具有固有活性的IF-1肿瘤，同时也产生关于HIF-1如何？稳定性受CDK的调节。